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G. Vattemi, G. Tomelleri, M. Filosto, C. Savio, N. Rizzuto and P. Tonin (2004)Neuropathology and Applied Neurobiology, doi:10.1111/j.1365-2990.2004.00602.xExpression of late myogenic differentiation markers in sarcoplasmic masses of patients with myotonic dystrophySarcoplasmic masses contain disorganized myofibrillar material and are a striking feature of myotonic dystrophy. However their significance is still unclear. Using immunocytochemistry we studied the expression of cytoskeletal proteins (desmin and vimentin), dystrophin, markers of myogenic differentiation (foetal myosin, neural cell adhesion molecule, bcl-2, insulin-like growth factor-I, fibroblast growth factor, retinoblastoma protein and myoD1), cell cycle regulators (Cdk2, p16, p27 and p57) and muscle proteases (ubiquitin,µ and m calpain and cathepsin D) in muscle biopsies from four patients with myotonic dystrophy. Sarcoplasmic masses were strongly positive for desmin, neural cell adhesion molecule, bcl-2, insulin-like growth factor I, retinoblastoma protein and p57, weakly positive for dystrophin and p16 and negative for vimentin, fibroblast growth factor, myoD1, Cdk2 and p27. Immunoreactivity for foetal myosin was detected only in a few fibres (< 1%). Our data suggest that the late myogenic differentiation programme is activated in sarcoplasmic masses although these areas do not reach complete maturation. [ABSTRACT FROM AUTHOR]

Summary A combined system for far- and near-field optical spectroscopy consisting of a compact scanning near-field optical microscope and a dedicated spectrometer was realized. The set-up allows the optical investigation of samples at temperatures from 10 to 300 K. The sample positioning range is as large as 5 × 5 × 5 mm3 and the spatial resolution is in the range of 1.5 µm in the far-field optical microscopy mode at low temperatures. In the scanning near-field optical microscope mode the resolution is defined by the microfabricated cantilever probe, which is placed in the focus of a double-mirror objective. The tip-to-sample distance in the scanning near-field optical microscope is controlled by a beam deflection system in dynamic scanning force microscopy mode. After a description of the apparatus, scanning force topography images of self-assembled InAs quantum dots on a GaAs substrate with a density of less than one dot per square micrometre are shown, followed by the first spectroscopic investigations of such a sample. The presented results demonstrate the potential of the system. [ABSTRACT FROM AUTHOR]

M. Filosto, P. Tonin, G. Vattemi, C. Savio, N. Rizzuto and G. Tomelleri (2003) Neuropathology and Applied Neurobiology 29, 52–59 Transcription factors c-Jun/activator protein-1 and nuclear factor-kappa B in oxidative stress response in mitochondrial diseases Mitochondrial dysfunction leads to oxygen free radical (ROS) generation with consequent oxidative stress and cellular damage. Recently, activation of the cellular antioxidant system and apoptosis were demonstrated in skeletal muscle fibres from patients with mitochondrial diseases, but the underlying mechanisms remain unknown. Hydrogen peroxide, a by-product of ROS generation, is a chemical inducer of gene expression able to activate apoptosis and to promote the antioxidant response through the activation of nuclear factor-kappa B (NF-κB) and activator protein-1 (AP-1) transcription factor. Using immunohistochemistry and confocal microscopy, we evaluated the expression of NF-κB and AP-1 in muscle biopsies from patients with mitochondrial disease. In addition, we examined the expression of factors involved in their activation, such as NF-κB inducing kinase (NIK) and phosphorylated Jun-N-terminal kinase (p-JNK). Most fibres with respiratory chain dysfunction displayed nuclear staining for activated c-Jun/AP-1, but not for NF-κB. The same fibres reacted for p-JNK. Only some ragged red fibres immunoreacted for NIK. These data suggest that AP-1 is involved in the oxidative stress response in muscle fibres from patients with mitochondrial disease. [ABSTRACT FROM AUTHOR]

We screened ATM gene mutations in 104 Italian Ataxia-Telangiectasia patients from 91 unrelated families (detection rate 90%) and found 21 recurrent mutations in 63 families. The majority (67%) of patients were compound heterozygotes, while 33% were homozygotes. To determine the existence of common haplotypes and potential founder effects, we analyzed five microsatellite markers within and flanking the ATM gene. Haplotype analysis was carried out in 48/63 families harbouring 16 of the 21 recurrent mutations. Forty different haplotypes were detected in the 48 A-T families studied. We found that the majority of patients with the same recurrent mutation originated from the same geographical area. All but one recurrent mutation analyzed displayed a common haplotype suggesting a single origin that then spread to different geographical areas. The high number of different haplotypes does not allow the screening of ATM mutations by haplotype analysis alone in the Italian population. The finding of recurrent public mutations without founder effect suggests the existence of 'mild' hot spots of mutation located along the sequence of the ATM gene.